Developing a standardized system of exposure and intervention endpoints for isoflurane in preclinical stroke models

Isoflurane is a regularly used anesthetic in translational research. Isoflurane facilitates invasive surgery and a rapid recovery. Specifically, in the pathology of stroke, controversy has surrounded isoflurane's intrinsic neuroprotective abilities, affecting apoptosis, excitotoxicity, and blood brain barrier disruption. Due to the intrinsic neuroprotective nature and lack of standardized guidelines for the use of isoflurane, research has shifted away from this gas in most animal models. Antagonistically, studies have also reported that no neuroprotective effects are observed when a surgery is accompanied with isoflurane exposure under 20 minutes. Isoflurane affects the pathophysiology in stroke patients by altering critical pathways in endothelial, neuronal, and microglial cells. Current studies have elucidated isoflurane neuroprotection to be time dependent and may be minimized in experimental designs if the exposure time is limited to a specific window. Therefore, with detailed and extensive literature on anesthetics, we can hypothesize that isoflurane exposure under the 20-minute benchmark, behavior and molecular pathways can be evaluated at any time-point following ischemic insult without confounding artifacts from isoflurane; however, If the exposure to isoflurane exceeds 20 minutes, the acute neuroprotective effects are evident for 2 weeks in the model, which should be accounted for in molecular and behavioral assessments, with either isoflurane inhibitors or a control group at 2 weeks post middle cerebral artery occlusion. The purpose of this review is to suggest a detailed and standardized outline for interventions and behavioral assessments after the use of isoflurane in experimental designs.